RE: Artifact: poor fixation or inadequate slide drying

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From:"Weems, Joyce" <>
To:'Adrienne Vair' <>,

If you coverslip from a xylene substitute - you might try using xylene
again. That stopped the identical complaints of our pathologists. Good luck!

> -----Original Message-----
> From:	Adrienne Vair []
> Sent:	Monday, May 29, 2000 1:57 AM
> To:
> Subject:	Artifact: poor fixation or inadequate slide drying
> Our pathologist's are concerned with the poor nuclear morphology they have
> been routinely seeing for many years; foamy appearance, muddy nuclear 
> staining (H&E).  I suggested that we look at extending the time in 10% 
> formalin as up until then, surgical specimens were received throughout the
> day from the O.R. in 10% Neutral Buffered Formalin (NBF) and most were
> being 
> processed the same day with the exception of bowel, uterus, and breast 
> tissue which were being opened and fixed 24 hours. We extented the
> fixation 
> time to a minimum of 24 hours for all surgical specimens including 
> colposcopy, endoscopy and fine needle biopsies but to no avail (and the 
> increase in turn around time concerned the physicians). The morphology 
> remained poor. I looked at temperatures of all reagents during processing 
> and embedding, and checked the pH of the 10% formalin being made up and
> used 
> routinely. I stopped using the microwave oven to dry slides as I could not
> find any literature to confirm that microwaving a rack of 20 slides for 7 
> minutes at full power (with small containers of water in the oven chamber)
> did not affect the tertiary structure of the proteins in the tissues. Nor 
> could I find a procedure in the lab or references as to where the idea 
> originated.  They are now being dried at 60 degrees celcius for one hour
> in 
> an oven prior to staining.  The pathologist's insisted that the problem
> was 
> not with fixation but with staining, I made improvements there but as I've
> stated before I could not improve on the nuclear morphology. My question
> is, 
> is this a fixation problem? Is this a slide drying problem as I have 
> recently read on this site?  I have read the article on alcohol/xylene 
> tissue processing (Journal of Histotechnology March 2000 issue page 45,
> and 
> the ideas seemed feasible but because it is unconventional, I would 
> appreciate some other ideas and comments.
> Thanks for the input.
> Adrienne Vair MLT BMLSc
> Histology Supervisor
> Medicine Hat Regional Hospital
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